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Objective To observe the effects of A2a adenosine receptor antagonist SCH442416 and ZM241385 on the expression of glutamine synthetase(GS) and L-Glutamate/L-Aspartate Transporter(GLAST) in rat retina under chronic ocular hypertension model.Methods Rat chronic ocular hypertension models were induced in the right eye of 12 male Sprague Dawley rats by blocking three episcleral veins,the left eye as control one.Intraocular pressure (IOP) was measured and compared at postoperative 1 week,2 weeks and 3 weeks.54 male chronic ocular hypertension rats were divided into 3 groups randomly,topically applying A2a adenosine receptor antagonist SCH442416,ZM241385 and carrier,respectively,three times a day for three weeks.At three weeks,mRNA and protein expression of GS and GLAST in rat retina were analyzed by RealTime-PCR and Western-blot.Results The average IOP of the modeling eyes at postoperative 1 week,2 weeks and 3 weeks were higher than that of the control eyes (all P ＜ 0.05).The mRNA and protein expression of GS and GLAST in the retina of SCH442416 and ZM241385 groups increased significantly compared to the carrier group (all P ＜ 0.05).However,the differences of mRNA and protein expression of GS and GLAST between SCH442416 and ZM241385 groups was not significant(all P ＞ 0.05).Conclusion Rat chronic ocular hypertension model can be induced by blocking three episceral veins successfully and effectively.A2a adenosine receptor antagonist SCH442416 and ZM241385 increase the expression of GS and GLAST.There seems no difference between the effects of these two drugs.
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The aim of this study was to determine the raising anticancer effects of resveratrol (Res) on paclitaxel (PA) in non-small cell lung cancer (NSCLC) cell line A549. The 10 µg/ml of Res had no effect on human fetal lung fibroblast MRC-5 cells or on A549 cancer cells and the 5 or 10 µg/ml of PA also had no effect on MRC-5 normal cells. PA-L (5 µg/ml) and PA-H (10 µg/ml) had the growth inhibitory effects in NSCLC cell line A549, and Res increased these growth inhibitory effects. By flow cytometry experiment, after Res (5 µg/ml)+PA-H (10 µg/ml) treatment, the A549 cells showed the most apoptosic cells compared to other group treatments, and after additional treatment with Res, the apoptosic cells of both two PA concentrations were raised. Res+PA could reduce the mRNA and protein expressions of COX-2, and Res+PA could reduce the COX-2 related genes of VEGF, MMP-1, MMP-2, MMP-9, NF-κB, Bcl-2, Bcl-xL, procollagen I, collagen I, collagen III and CTGF, TNF-α, IL-1β, iNOS and raise the TIMP-1, TIMP-2, TIMP-3, IκB-α, p53, p21, caspase-3, caspase-8, caspase-9, Bax genes compared to the control cells and the PA treated cells. From these results, it can be suggested that Res could raise the anticancer effects of PA in A549 cells, thus Res might be used as a good sensitizing agent for PA.
Subject(s)
Humans , Carcinoma, Non-Small-Cell Lung , Caspase 3 , Caspase 8 , Caspase 9 , Cell Line , Collagen , Fibroblasts , Flow Cytometry , In Vitro Techniques , Lung , Paclitaxel , Procollagen , RNA, Messenger , Tissue Inhibitor of Metalloproteinase-1 , Tissue Inhibitor of Metalloproteinase-2 , Tissue Inhibitor of Metalloproteinase-3 , Vascular Endothelial Growth Factor AABSTRACT
Based on the construction and management practice of the morphologic experimental center in Xi'an Medical University, the achievements in laboratory daily operation and institutional man-agement were summarized in the area of lab rules and regulations, instrument and equipment, experiment teaching, lab environment and safety, lab staff administration and so on. The management work has been refined using the practice model of resource sharing, system administration, individual responsibility, and unified staff supervision. The lab rules and responsibilities were also effectively implemented on specific person. Taking the opportunity in teaching evaluation at the experimental center, the lab connotation con-struction was further strengthened. The evaluation system was thoroughly examined in order to look for gaps and promote the lab construction. Further work could be carried out on the laboratory software and hard-ware, such as instrument and equipment update, experimental teaching system reformation in morphology, promotion on multidisciplinary integration and unified management of lab staff.
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Purpose To observe the mutation of K-ras gene and expression of Fascin-1 protein in CRC tissues and their relationship with clinical pathological features, and then to analyze the correlation between mutation of K-ras and expression of Fascin-1. Methods In 86 cases of CRC tissues, K-ras mutation was detected by DNA sequencing analysis, and Fascin-1 expression was detected by im-munohistochemical method. Results In CRC tissues the mutation rate of K-ras was 34. 88%, the expression rate of Fascin-1 was 60. 47%. The mutation rate of K-ras in lymph node metastasis group was higher than that of without lymph node metastasis group, and that in distant metastasis group was higher than that of without distant metastasis group(P<0. 05). The expression rate of Fascin-1 in serosa invasion group was higher than that of without serosa invasion group, and that in lymph node metastasis group was higher than that of without lymph node metastasis group, and that in distant metastasis group was higher than that of without distant metastasis group (P<0. 01). There was a correlation between the mutation of K-ras gene and the expression of Fascin-1 in CRC tissues (rp =0. 236, P<0. 05). Conclusions The CRC tissues with mutation of K-ras are more likely to metastasize and the CRC tissues with expression of Fascin-1 are more likely to invade serosa and metastasize. The CRC tissues with mutation of K-ras are more likely to express Fascin-1.
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BACKGROUND: Excellent Iow-antigenicity xenogeneic biological valve scaffold is the premise of constructing tissue-engineered valve by using which kind of acellular methods.OBJECTIVE: To explore the optimal preparation method of making tissue engineered heart valves by meesuring efficiency of different acellular methods and ability to preserve the matrix.DESIGN, TIME AND SETTING: The prospective randomly controlled study was performed at the Central Laboratory of Taian Central Hospital from January 2007 to June 2008.MATERIALS: Sixteen specimens of porcine aortic valves were randomly divided into control, NaCI-sodium-dodecyl-sulfate (SDS),trypsin and triton-X100 groups.METHODS: Specimens in the control group were left intact. Three test groups were decelluladzed with NaCI, trypsin andTriton-X100 respectively.MAIN OUTCOME MEASURES: The gross structure, optical and electron microscope ultrastructure of the decelluarated porcineheart valve matrix was compared. The expression of vascular endothelial cell major histocompatibility complex (MHC)- Ⅰ antigenwas detected by immunohistochemical method.RESULTS: Treatment with NaCL-SDS achieved only incomplete decellularization. The main components of extracellular matdxwere reserved completely, but the fibrous components became unclear and swelling. Treatment with trypsin removed cellscompletely, but caused serious structural alterations, with the presence of swollen collagen fiber, crude edge, widen and irregularfiber interspace. Treatment with Triton-X100 achieved both complete decelluarization and preservation of the matrix structure.Valves following treatment of NaCI-SDS, trypsin and Triton-X100 had certain immunogenicity. However, the immunogenicity ofvalves following treatment of trypsin and Triton-X100 was significantly lower compared with the treatment of NaCL-SDS.CONCLUSION: The decellularization method by Triton-X100 is effective and complete. The Triton-X100 method does not changematrix structure and has low immunogenicity.
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Objective To investigate the effect of TGIF on apoptosis of TGF-?-induced gastric canceer cell.Methods After TGIF was stably transfected into gastric cancer cell line BGC823,apoptosis was examined with flow cytometry,and the expressions of TGIF,caspase8 and caspase9 were analyzed with Western blot.Results The apoptosis rate of BGC823 cells with the treatment of TGF-? obviously increased,and was accompanied by activation of caspase9 but not caspase8.Moreover,compared with controls,pcDNA3.1-TGIF-transfected BGC823 cells significantly decreased the sensitivity of TGF-?-induced apoptosis.Conclusions TGF-? induced the apoptosis of gastric cancer cell BGC823 via caspase9 pathway;furthermore,TGIF could inhibit TGF-?-induced apoptosis.